Effect of extraction time and acid concentration on the separation of proglycogen and macroglycogen in horse muscle samples.

The objective of this study was to determine whether the concentrations of proglycogen (PG) and macroglycogen (MG) in biopsy samples of horse muscle are influenced by extraction time or perchloric acid (PCA) concentration. In study 1, individual muscle-biopsy samples from 10 horses were divided into 4 parts each and then randomly subjected to 4 periods of extraction (10, 20, 60, or 120 min) with 1.5 M PCA. In study 2, individual muscle-biopsy samples from 6 horses were divided into 24 pieces each and then randomly subjected to 12 combinations of extraction time (10, 20, 30, or 40 min) and PCA concentration (0.5, 1.5, or 3.0 M). The results from study 1 indicated that PG and MG concentrations are affected only after extraction for 120 min; the PG concentration decreased significantly (P < 0.05), and the MG concentration increased (not significantly). In study 2, extraction in 3.0 M PCA yielded significantly lower PG and higher MG concentrations (P < 0.05) than extraction in 0.5 or 1.5 M PCA with each of the extraction times. The results of this study further support the existence of 2 glycogen pools and demonstrate that they are not an extraction artifact. The study also suggests that the 2 pools are stable during extraction over a range of extraction times and acid concentrations. However, if the exposure to acid is very long and, or, the acid concentration is high, some of the insoluble PG appears to be hydrolyzed and to enter the MG pool.